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You are here:Home » Cloning » Cloning-Enzymes » Ribonuclease H (RNase H)

Ribonuclease H (RNase H)

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Specifications

Ribonuclease H specifically degrades only the RNA strand in RNA-DNA hybrids. It does not hydrolyze the phosphodiester bonds within single-stranded or double-stranded DNA and RNA.
Catalog #R2021
Unit DefinitionOne unit catalyzes the formation of 1nmol of acid soluble products in 20min at 37ºC. Enzyme activity is assayed in the following mixture: 20mM Tris-HCl (H 7.8), 40 mM KCl, 8mM MgCl2, 1mM DTT, 24uM [3H]-POLY(A)-POLY(dT), 0.03mg/ml BSA AND 4% Glycerol.
10X Reaction buffer (included)R2021A
Supplied as a liquid in 200mM Tris HCl, pH 8.0, 400mM KCl, 80mM MgCl2, 10mM DTT.
ApplicationsSuitable for use in Removal of mRNA prior to the synthesis of the 2nd strand of cDNA, Removal of mRNA poly(A) sequences after hybridization with oligo(dT), Site-specific cleavage of RNA and Studies of in vitro polyadenylation rection products.
Inhibition and InactivationInhibitors: metal chelators, SH-blocking reagents.
-Inactivated by heating at 65ºC for 10min.
Endodeoxyribonuclease AssayNo detectable conversion of covalently closed circular DNA to nicked DNA was observed after incubation of 10 units of Ribonuclease H with 1ug of pBR322 DNA
in 50ul of reaction buffer for 1 hour at 37°C.
Exodeoxyribonuclease Assay0% of the total radioactivity was released into trichloroacetic acid-soluble fraction after incubation of 10 units of Ribonuclease H with 1ug of sonicated
E.coli [3H]-DNA in 50ul of reaction buffer for 1 hour at 37°C.
Ribonuclease Assay0% of the total radioactivity was released into trichloroacetic acid-soluble fraction after incubation of 10 units of Ribonuclease H with 1ug of [3H]-RNA in 50ul of reaction buffer for 1 hour at 37°C.
Storage and StabilityFor long-term storage, aliquot and store at -20°C. Aliquots are stable for at least 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
CAS Number9050-76-4
Molecular Weight19kD
SourceE. coli MRE-600 cells
Concentration~5u/ul
FormSupplied as a liquid in 25mM HEPES-KOH, pH 8.0, 50mM KCl, 1mM DTT, 1mM EDTA, 0.1mg/ml BSA, and 50% glycerol.
Important NoteThis product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
Alternate namesE. coli; cRNAse H


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